title: Single-molecule force spectroscopy of G-protein-coupled receptors.
abstract: The applicability of single-molecule force spectroscopy (SMFS) to characterize membrane proteins in vitro is developing rapidly and opening a wide range of fascinating possibilities to study how intra- and intermolecular interactions determine their structural stability and functional state. In particular, understanding how molecular interactions contribute to the functional state of G-protein-coupled receptors (GPCRs) is of importance because they mediate most of our physiological responses and act as therapeutic targets for a broad spectrum of diseases. In our review we focus on SMFS to characterize GPCRs embedded in their physiologically relevant membranes and exposed to physiologically relevant conditions. SMFS uses a molecularly sharp stylus to grasp the terminal end of a GPCR and to quickly unfold the receptor while recording interaction forces. The positional accuracy of SMFS localizes these interactions to structural segments of the GPCR whereas the sensitivity of SMFS enables their stabilizing interaction forces to be quantified. To further investigate the kinetic, energetic and mechanical properties of the structural segments, dynamic SMFS (DFS) probes their stability over a wide range of loading rates. These parameters provide insight into the energy landscape that provides information on the structural and functional properties of the GPCRs. Selected highlights exemplify the application of SMFS to characterize inter- and intramolecular interactions, which change the properties of GPCRs in relation to their functional state (e.g., ligand binding), diseased state (e.g., mutation), or lipid environment such as cholesterol.
authors.affiliation: Department of Biosystems Science and Engineering, ETH Zürich, 4058 Basel, Switzerland. daniel.mueller@bsse.ethz.ch.
authors.forename: Michael
authors.initials: M
authors.lastname: Zocher
authors.forename: Christian A
authors.initials: CA
authors.lastname: Bippes
authors.forename: Cheng
authors.initials: C
authors.lastname: Zhang
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Müller
mesh_terms: D000818:Animals; D002417:Cattle; D006801:Humans; D018625:Microscopy, Atomic Force; D011499:Protein Processing, Post-Translational; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry*; D012243:Rhodopsin / Q000737:chemistry
chemical_list: D043562:Receptors, G-Protein-Coupled; D012243:Rhodopsin